Figure 3 .

STIM2 does not regulate AMPK-mTORC1 signaling upon energy stress. A) Glucose starvation does not alter AMPK binding to STIM2. Cells stably expressing STIM2-SFB were cultured in glucose-containing and glucose-free medium for 6 h and subjected for immunoprecipitation by S-protein beads. Coprecipitated proteins were analyzed after Western blotting. B, C) STIM2 levels in 786-O (B) and HeLa (C) cells infected with control sgRNA (Ctl sg), STIM2 sgRNA-1 (STIM2 sg1), and STIM2 sgRNA-2 (STIM2 sg2) were measured by Western blotting. D, E) Energy stress does not alter AMPK and mTORC1 activation in STIM2-depleted cells. STIM2-deficient 786-O cells were cultured in glucose-containing or glucose-free medium for 24 h (D) or treated with 2 deoxy-glucose (5 mM) for 18 h (E). Cell lysates were analyzed by immunoblotting for the phosphorylated and total levels of indicated proteins. F–I) STIM2 depletion does not affect glucose starvation–induced cell death. STIM2-deficient 786-O (F, H) and HeLa (G, I) cells were cultured in glucose-containing or glucose-free medium for 48 or 6 h, respectively. Representative images (F, G) and quantification of the cell death (H, I) were analyzed after glucose starvation. Ns, not significant. P > 0.05.